Fraunhofer Institut für Molekularbiologie und Angewandte Okologie, Forckenbeckstrasse 6, 52074 Aachen, Germany.
Appl Microbiol Biotechnol. 2010 Oct;88(3):689-98. doi: 10.1007/s00253-010-2771-4. Epub 2010 Jul 28.
Forisomes are mechanoproteins that undergo ATP-independent contraction-expansion cycles triggered by divalent cations, pH changes, and electrical stimuli. Although native forisomes from Medicago truncatula comprise a number of subunits encoded by separate genes, here we show that at least two of those subunits (MtSEO1 and MtSEO4) can assemble into homomeric forisome bodies that are functionally similar to their native, multimeric counterparts. We expressed these subunits in plants and yeast, resulting in the purification of large quantities of artificial forisomes with unique characteristics depending on the expression platform. These artificial forisomes were able to contract and expand in vitro like native forisomes and could respond to electrical stimulation when immobilized between interdigital transducer electrodes. These results indicate that recombinant artificial forisomes with specific characteristics can be prepared in large amounts and used as components of microscale and nanoscale devices.
成孔蛋白是机械蛋白,可在二价阳离子、pH 值变化和电刺激的作用下发生无需 ATP 的收缩-舒张循环。尽管来自紫花苜蓿的天然成孔蛋白由许多由独立基因编码的亚基组成,但在这里我们表明,其中至少两个亚基(MtSEO1 和 MtSEO4)可以组装成同源成孔体,其功能与天然的多聚体成孔体相似。我们在植物和酵母中表达了这些亚基,从而大量纯化了具有独特特性的人工成孔体,具体特性取决于表达平台。这些人工成孔体在体外能够像天然成孔体一样收缩和扩张,并且当固定在叉指换能器电极之间时能够对电刺激做出反应。这些结果表明,可以大量制备具有特定特性的重组人工成孔体,并将其用作微尺度和纳米尺度装置的组件。